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Background: Odorant binding proteins (OBPs) initiate the process of odorant perception. Numerous investigations have demonstrated that OBPs bind a broad variety of chemicals and are more likely to carry pheromones or odor molecules with high binding affinities. However, few studies have investigated its effects on insect behavior. Previously, we found that AmelOBP4 has a significantly higher expression in the heads of foragers than that of nurses regardless of their ages, revealing its importance in foraging behaviour of the honey bee. RNA interference (RNAi) is the induction of sequence specific gene silencing by double-stranded RNA (dsRNA), it is a powerful tool that makes gene inactivation possible in organisms that were not amenable to genetic analysis before. Results: In this study, we found that AmelOBP4 had high expression levels in the antennae of both nurses and foragers, and could be successfully inhibited by feeding double stranded RNA of AmelOBP4 (dsAmelOBP4). Foragers with inhibited AmelOBP4 showed significantly lower sugar responsiveness than control bees, and also significantly reduced EAG response to plant volatiles of nonanal, linalool and 1-Octen-3ol. On the other hand, nurses with inhibited AmelOBP4 showed significantly reduced EAG response to brood pheromone of ethyl oleate, methyl linoleate, methyl palmitate and β-ocimene. Finally, the Y-tube choice assay showed nurses only exhibited a significantly reduced preference to ethyl oleate, but foragers exhibited significantly reduced preference to all these three plant volatiles. Conclusions: The findings of our study suggested that AmelOBP4 plays an important role in the odorant binding process, especially in modulating olfactory behaviour in workers. Our results provide a foundation for exploring the olfactory mechanism of Apis mellifera. [ABSTRACT FROM AUTHOR]

BACKGROUND: The parasitic mite, Varroa destructor has posed a threat to the health and survival of European honey bees, Apis mellifera worldwide. There is a prevailing belief that small comb cells could provide a management tool against Varroa mites. However, the hypothesis that smaller cells can impede Varroa reproduction has not been fully tested. Here, we tested this hypothesis under laboratory conditions by using two distinct Varroa in vitro rearing systems: one involved gelatin capsules of different sizes, specifically size 00 (0.95 mL) versus size 1 (0.48 mL), and the second consisted of brood comb cells drawn on 3D printed foundations with varying cell sizes, ranging from 5.0 mm to 7.0 mm at 0.5 mm intervals. RESULTS: The results showed that mother mites in size 00 cells had significantly lower fecundity and fertility compared to those in size 1 cells. Interestingly, the reproductive suppression in larger cells could be reversed by adding an extra worker larva. Similarly, gonopore size of mother mites was smaller in size 00 cells, but restored with another host larva. Furthermore, both the fecundity and fertility of mother mites decreased linearly with the size of brood comb cells. CONCLUSIONS: Our results suggest that the reproduction of V. destructor is hindered by larger cells, possibly because larger brood cells disperse or weaken host volatile chemical cues that are crucial for Varroa reproduction. The insights derived from this study are expected to hold significant implications for the implementation of Varroa management programs. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Variation in endocrine signaling is proposed to underlie the evolution and regulation of social life histories, but the genetic architecture of endocrine signaling is still poorly understood. An excellent example of a hormonally influenced set of social traits is found in the honey bee (Apis mellifera): a dynamic and mutually suppressive relationship between juvenile hormone (JH) and the yolk precursor protein vitellogenin (Vg) regulates behavioral maturation and foraging of workers. Several other traits cosegregate with these behavioral phenotypes, comprising the pollen hoarding syndrome (PHS) one of the best-described animal behavioral syndromes. Genotype differences in responsiveness of JH to Vg are a potential mechanistic basis for the PHS. Here, we reduced Vg expression via RNA interference in progeny from a backcross between 2 selected lines of honey bees that differ in JH responsiveness to Vg reduction and measured JH response and ovary size, which represents another key aspect of the PHS. Genetic mapping based on restriction site-associated DNA tag sequencing identified suggestive quantitative trait loci (QTL) for ovary size and JH responsiveness. We confirmed genetic effects on both traits near many QTL that had been identified previously for their effect on various PHS traits. Thus, our results support a role for endocrine control of complex traits at a genetic level. Furthermore, this first example of a genetic map of a hormonal response to gene knockdown in a social insect helps to refine the genetic understanding of complex behaviors and the physiology that may underlie behavioral control in general.

The Western honey bee (Apis mellifera) is a social insect characterized by caste differentiation in which the queen bee and worker bees display marked differences in morphology, behavior, reproduction, and longevity despite their identical genomes. The main causative factor in caste differentiation is the food fed to queen larvae, termed royal jelly (RJ). Alternative splicing (AS) is an important RNA-mediated post-transcriptional process in eukaryotes. Here we report AS changes in A. mellifera after being fed either A. mellifera RJ or A. cerana RJ. The results demonstrated that the RJ type affected 4 types of AS in adult A. mellifera: exon skipping, intron retention, alternative 5’ splice sites, and alternative 3’splice sites. After feeding with A. cerana RJ, AS occurred in many genes in adult A. mellifera that encode proteins involved in development, growth, the tricarboxylic acid cycle, and substance metabolism. This study provides the first evidence that heterospecific RJ can influence the AS of many genes related to honey bee development and growth.

Removing dead conspecifics reduces pathogen transmission in social insects. Undertakers of the hive-bees can quickly detect and remove dead bees. The signals indicating instant death are unclear. In this study, we identified that undertaking was quickly elicited by the reduction in evaporated cuticular hydrocarbons (CHC emissions, CHEs). CHEs from dead bees were much lower than those from live bees, whereas heated dead bees with reproduced CHEs were not instantly removed. Physiological tests further showed that undertakers perceived and discriminated the major CHCs, and heated synthetic CHCs inhibited the undertaking behaviour. This study thus indicates that the reduced emissions of CHCs, particularly heptacosane and nonacosane, due to lower body temperatures in dead bees, are used by undertakers as a signal for detecting dead bees. Heptacosane and nonacosane emissions at hive temperatures are life signals. By changing the vapour pressure, then the ratio of emitted compounds, insect chemical communication can be fine-tuned by body temperature. The increasing frequency of extreme weather events may cause inaccurate death recognition, harming bee health. [ABSTRACT FROM AUTHOR]

Simple Summary: Honey bees provide essential pollination services for many agricultural crops and wild plants. However, honey bee health is at risk. As an obligate ectoparasite of honey bees, Varroa destructor has been causing huge damage to the worldwide Western honey bee colonies since the host shift from its original host, the Eastern honey bees. For years, a lot of effort has been made to control this parasite; however, more efficient and safer methods are still urgently needed. The current study also aimed to contribute to the Varroa mitigation by seeking the potential compounds that affected the host-finding behavior of Varroa mites. By profiling the headspace volatiles of honey bee larvae, four aliphatic esters were commonly identified from both worker and drone larvae of the original and new hosts. Among the esters, ethyl myristate was a novel compound and able to significantly attract the Varroa mites. The findings presented in this paper will deepen our understanding of the Varroa host finding behavior and provide new chemicals for the monitoring and control of V. destructor in the future. Varroa destructor is an obligate ectoparasite of honey bees and shifted from its original host Apis cerana to the new host Apis mellifera in the first half of the twentieth century. The host shift has resulted in a great threat to the health and survival of A. mellifera colonies worldwide. Chemical signals play a crucial role in all aspects of the Varroa life cycle, including host finding. However, the chemical cues that affect the host finding behavior of Varroa mites are still not fully understood. In this study, we systematically profiled the headspace volatiles of both worker and drone larvae of the two honey bee species by using solid phase micro-extraction coupled to gas chromatography-mass spectrometry (SPME-GC-MS), and then used electrophysiological recording and Y-tube olfactometer bioassay to study the potential roles of the selected compounds. The chemical profiling showed that there were four aliphatic esters, ethyl myristate (EM), methyl palmitate (MP), ethyl palmitate (EP), and ethyl oleate (EO) commonly detected from all four types of larval hosts. Among them, EM was a new substance identified from honey bee headspace volatiles. Results from electrophysiological recordings indicated that all the aliphatic esters could elicit significant responses of Varroa pit organs on its forelegs. Moreover, behavioral analyses revealed that EM could significantly attract V. destructor at a medium dosage (10 µg), while MP had no observable effect on the mites and both EP and EO were able to repel the parasites. Our findings suggest an important role of host-derived aliphatic esters in Varroa host finding, and provide new chemicals for Varroa monitoring and control. [ABSTRACT FROM AUTHOR]

Sex-determination mechanisms vary greatly among taxa. It has been proposed that genetic sex-determination pathways evolve in reverse order from the final step in the pathway to the first step. Consistent with this hypothesis, doublesex (dsx), the most downstream gene in the Drosophila sex-determination cascade that determines most sexual phenotypes also determines sex in other dipterans and the silk moth, while the upstream genes vary among these species. However, it is unknown when dsx was recruited to the sex-determination pathway during insect evolution. Furthermore, sex-specific splicing of dsx, by which dsx determines sex, is different in pattern and mechanism between the moth and the fly, raising an interesting question of how these insects have kept the executor of sex determination while allowing flexibility in the means of execution. To address these questions, here we study the dsx gene of the honeybee Apis mellifera, a member of the most basal lineage of holometabolous insects. We report that honeybee dsx is sex-specifically spliced and that it produces both the fly-type and moth-type splicing forms, indicating that the use of different splicing forms of Dsx in controlling sexual differentiation was present in the common ancestor of holometabolous insects. Our data suggest that in ancestral holometabolous insects the female Dsx form is the default and the male form is generated by suppressing the splicing of the female form. Thus, it is likely that the dsx splicing activator system in flies, where the male form is the default, arose during early dipteran evolution.

Brain transcriptional regulatory network for behavior demonstrates that brain gene expression in the honey bee can be accurately predicted from the expression transcription factors (TFs), but roles for specific TFs are less understood. Mushroom bodies (MBs) are important for learning, memory and sensory integration in the honey bee brain. A TFs, Mblk‐1, expressed preferentially in the large‐type Kenyon cells of the honeybee MBs is predicted to be involved in brain function by regulating transcription of its target genes in honey bee. However, its function and the mechanism of regulation in behavior of honey bee is still obscure. Here we show that Mblk‐1 had significantly higher expression in the brains of forager bees relative to nurse bees. Mblk‐1 was significantly inhibited in bees fed small interfering RNA. In addition, inhibition of Mblk‐1 decreased sucrose responsiveness in foragers. Finally, we determined that Mblk‐1 regulated the messenger RNA of AmGR1. These findings suggest that Mblk‐1 may target AmGR1 to regulate the sucrose responsiveness of foragers. [ABSTRACT FROM AUTHOR]